| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 565/575 |
| MW | N/A |
| CAS# | N/A |
| Solvent | Water |
| Storage | R/D/L |
| Category | Protein-BasedTags Phycoproteins |
| Related | ProteinDyes |
| Spectrum | AdvancedSpectrumViewer |
- Pre-activateAntibodywithBuccutite™MTA
- ReconstituteBuccutite™MTAinDMSOat~10mg/mL.
Note:PleasestoreunusedMTAat-20°Candcouldbeuseduptotwofreezeandthawcycles. - Preparetargetantibody(Ab)inpH=8.5~9.0bufferatconcentrationabove1mg/ml
- AddtheMTAtoAbsolutionattheratioof8~10µgMTA/100µgAb.
- MixwellandreactatRTfor60minutes,rotatingduringthereaction.
- PurifythereactionmixturewithdesaltingcolumntoremoveunreactedMTAandexchangebuffertoPBSorbufferofyourchoice.
- CollecttheMTA-activatedAb,andestimatetheconcentrationby70%yieldoftheoriginalstartingamount.
- ReconstituteBuccutite™MTAinDMSOat~10mg/mL.
- ConjugatewithPre-activatedPE,PE-iFluors,orPE-iFluorTandems
- Reconstitutepre-activatedPE,PE,PE-iFluors,orPE-iFluorTandemsin100µLddH2Oto10mg/mL.
Note:Reconstitutedpre-activatedPE,PE-iFluors,orPE-iFluorTandemsarenotstableandcouldbestoredat4°Cforonemonth,pleasekeptitfromlight. - Addpre-activatedPE,PE-iFluors,orPE-iFluorTandemsdirectlytoMTA-activatedtargetAbsolution(from1.5)attheratioof300µgPE-iFluor647/100µgMTA-activatedAb.
- Rotatethemixturefor1~2hoursatroomtemperature
- TheAb/PE,PE-iFluors,orPE-iFluorTandemsconjugatesarenowreadytouse.
Optional:Ab/PE,PE-iFluors,orPE-iFluorTandemsconjugatecouldbefurtherpurifiedthroughsizeexclusionchromatographytogetbestperformance.
Note1:Theantibodyconjugateshouldbestoredat>0.5mg/mLinthepresenceofacarrierprotein(e.g.,0.1%bovineserumalbumin)and0.02-0.05%sodiumazide.
Note2:TheAb/PE,PE-iFluors,orPE-iFluorTandemsconjugatesolutioncouldbestoredat4°Cfortwomonthsandkeptfromlight.
- Reconstitutepre-activatedPE,PE,PE-iFluors,orPE-iFluorTandemsin100µLddH2Oto10mg/mL.
| References&Citations |  PrinterFriendlyVersion |
1. ZhaoKH,SuP,LiJ,TuJM,ZhouM,BubenzerC,ScheerH.(2006)Chromophoreattachmenttophycobiliproteinbeta-subunits:phycocyanobilin:cysteine-beta84phycobiliproteinlyaseactivityofCpeS-likeproteinfromAnabaenaSp.PCC7120.JBiolChem,281,8573.
2. PetrasekZ,SchmittFJ,TheissC,HuyerJ,ChenM,LarkumA,EichlerHJ,KemnitzK,EckertHJ.(2005)ExcitationenergytransferfromphycobiliproteintochlorophylldinintactcellsofAcaryochlorismarinastudiedbytime-andwavelength-resolvedfluorescencespectroscopy.PhotochemPhotobiolSci,4,1016.
3. LoosD,CotletM,DeSchryverF,HabuchiS,HofkensJ.(2004)Single-moleculespectroscopyselectivelyprobesdonorandacceptorchromophoresinthephycobiliproteinallophycocyanin.BiophysJ,87,2598.
4. PrasannaR,PrasannaBM,MohammADISA,SinghPK.(2003)EvaluationofTolypothrixgermplasmforphycobiliproteincontent.FoliaMicrobiol(Praha),48,59.
5. PrasannaR,DharDW,DominicTK,TiwariON,SinghPK.(2003)IsolationandcharacterisationofphycobiliproteinrichmutantofcyanobacteriumSynechocystissp.ActaBiolHung,54,113.
6. WuP.(2000)[Phycobiliproteinandfluorescenceimmunologicalassay].ShengLiKeXueJinZhan,31,82.
7. NoubirS,LuqueI,OchoadeAldaJA,PerewoskaI,TandeaudeMarsacN,CobleyJG,HoumardJ.(2002)Co-ordinatedexpressionofphycobiliproteinoperonsinthechromaticallyadaptingcyanobacteriumCalothrixPCC7601:aroleforRcaDandRcaG.MolMicrobiol,43,749.
8. TingCS,RocapG,KingJ,ChisholmSW.(2001)PhycobiliproteingenesofthemarinephotosyntheticprokaryoteProchlorococcus:evidenceforrapidevolutionofgeneticheterogeneity.MicroBIOLOGy,147,3171.
9. TriantafilouK,TriantafilouM,WilsonKM.(2000)Phycobiliprotein-Fabconjugatesasprobesforsingleparticlefluorescenceimaging.Cytometry,41,226.
10. ZhaoKH,DengMG,ZhengM,ZhouM,ParbelA,StorfM,MeyerM,StrohmannB,ScheerH.(2000)Novelactivityofaphycobiliproteinlyase:boththeattachmentofphycocyanobilinandtheisomerizationtophycoviolobilinarecatalyzedbytheproteinsPecEandPecFencodedbythephycoerythrocyaninoperon.FEBSLett,469,9.
